Karyotypes of Capoeta antalyensis (Battalgil, 1944) and Capoeta baliki Turan, Kottelat, Ekmekçi & İmamoğlu, 2006 (Actinopterygii, Cyprinidae)

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1 Turkish Journal of Fisheries and Aquatic Sciences 17: (2017) ISSN DOI: / v17_2_05 RESEARCH PAPER Karyotypes of Capoeta antalyensis (Battalgil, 1944) and Capoeta baliki Turan, Kottelat, Ekmekçi & İmamoğlu, 2006 (Actinopterygii, Cyprinidae) Muradiye Karasu Ayata 1, Sevgi Ünal 2,*, Muhammet Gaffaroğlu 1 1 Department of Biology, Faculty of Science and Arts, Ahi Evran University, Kırşehir, Türkiye. 2 Department of Biology, Faculty of Science, Gazi University, Ankara, Türkiye. * Corresponding Author: Tel.: ; sevgiunal@ymail.com Received 15 March 2016 Accepted 08 September 2016 Abstract Chromosome numbers and morphologies of Capoeta antalyensis (Battalgil, 1944) originating from Boğa Creek and Capoeta baliki Turan, Kottelat, Ekmekçi & İmamoğlu, 2006 originating from Kızılırmak River were investigated. Four females and two males specimens of C. antalyensis and three females and five males specimens of C. baliki were analyzed. Metaphase chromosomes were obtained from kidney cells. The diploid chromosome number of C. antalyensis was found 2n=150, of which 42 pairs were meta-submetacentric chromosome and 33 pairs were subtelo-acrocentric chromosome, and fundamental arm number (NF) was found 234. The diploid chromosome number of C. baliki was 2n=150, consisting of 44 meta-submetacentric chromosome pairs and 31 subtelo-acrocentric chromosome pairs, and the number of arms was 238. Neither species showed any sex chromosome differentiation. Keywords: Capoeta antalyensis, Capoeta baliki, karyotype, Anatolia. Introductıon It is known that 19 species of the genus Capoeta Cuiver-Valenciennes 1842, belonging to Cyprinidae family live in the inland waters of Turkey. Capoeta antalyensis is an endemic species that prevails in the rivers in the vicinity of Antalya Province. C. baliki, previously was named as Capoeta tinca, is another endemic species that pervades in Sakarya and Kızılırmak Rivers (Geldiay and Balık, 2007; Kuru et al., 2014). Polyploidy as one of the most striking aspects of fish genetics can also be analyzed with chromosome counts (Thorgaard and Disney, 1990). In a study about the karyology of five Barbus species in South Africa, Oellerman and Skelton (1990) found that chromosome counts ranged between 2n=148 and 2n=150 with a majority of the species in the Cyprinidae family having 2n=50 chromosomes, and argued that the latter species were of hexaploid origin. Rab and Collares-Pereira (1995), on the other hand, stated that Barbus species were cyprinids of tetraploid origin and were characterized by 2n=100 diploid count. According to these authors, polyploidy in cyprinid fish is an extremely complicated event resulting from various origins and the chromosome number in polyploid species increases in integral Published by Central Fisheries Research Institute (CFRI) Trabzon, Turkey in cooperation w ith Japan International Cooperation Agency (JICA), Japan multiples of the most common chromosome value (2n=50). It was noted that Barbus bynni (Syn: Barbus bynni occidentalis) and B. wurtzi had a chromosome number of 2n=148 and B. petitjeani had a chromosome number of 2n=150 and that all three species were hexaploid (Guegan et al., 1995). Chromosome number and morphology can vary intra and interspecifically. Analysis of this variation within and among species is currently a popular approach which is widely used by fish systematists. While intraspecific variations can be used for analysis of population structure and dynamics, interspecific variations are useful sources to apply for analyzing an array of evolutionary and genetic hypotheses. For this purposes the research of fish chromosomes has become an important area (Thorgaard and Disney, 1990). Although many cytogenetic studies have been carried out on Anatolian fishes (Gaffaroğlu et al., 2006; Gaffaroglu et al., 2012) no cytogenetic study about C. antalyensis and C. baliki has been found. The present study is the first to examine the karyotype characteristics of C. antalyensis and C. baliki. Materials And Methods Specimens of C. antalyensis (four females and two males) originating from Boga Creek, Antalya,

2 270 M. K. Ayata et al. / Turk. J. Fish. Aquat. Sci. 17: (2017) Türkiye (36 51'N, 30 37'E) and C. baliki (three females and five males) originating from Kızılırmak River, Kırşehir, Türkiye (38 57'N, 34 12'E) were analyzed (Figure 1). They were transported alive to Discussion the laboratory and kept in well-aerated aquaria until analysis. Mitotic chromosome slides were prepared according to Collares-Pereira (1992) from kidney cells. The specimens were injected intraperitoneally with 0.1% colchicine solution and head kidneys of specimens were removed and placed in KCl solution. The cell suspension was centrifuged and supernatant was discarded. The cell suspensions were dropped onto cleaned slides. The slides were stained with 10% Giemsa. At least 10 metaphases were counted per specimen. Chromosomes were classified using the nomenclatures proposed by Levan et al. (1964). Metasubmetacentric (M-SM) chromosomes were taken as biarmed while subtelo-acrocentric (ST-A) chromosomes were taken as uniarmed. Classification of chromosomes was made according to ratio of long and short arm. Metacentric (M) means a chromosomes with equal-sized arms, Submetacentric (SM) means a chromosomes with the ratio of long arm more than the ratio of short arm. ST -A means a chromosomes with the short arm at the end of centromere and/or centromere is non-terminal (uniarmed). The preparations were observed and photographed digitally at a Leica DMLB 3000 research microscope. Results Diploid chromosome numbers of C. antalyensis and C. baliki were determined to be 2n=150. Chromosome morphology of C. antalyensis consisted of 42 pairs of M-SM and 33 pairs of subteloacrocentric ST-A chromosomes with NF 234 (Figure 2) and C. baliki had 44 pairs of M -SM and 31 pairs of ST-A chromosomes with NF 238 (Figure 3). There was no sex chromosome differentiation in these two species. A review of literature has shown that there is no previous cytogenetic study about C. antalyensis and C. baliki. The present study is the first to determine the chromosome number and morphology of C. antalyensis and C. baliki and to characterize their karyotype. Diploid chromosome numbers of C. antalyensis and C. baliki have been found identical. However, there are differences in their chromosome morphologies. Two pairs of chromosomes identified as ST-A in C. antalyensis were determined to be M- SM in C. baliki. Due to the differences in their chromosome morphologies, NF of C. antalyensis and C. baliki were also found different. Results obtained from C. antalyensis and C. baliki are similar to those found in other Anatolian Capoeta species (Table 1). Capoeta trutta and Capoeta umbla (Syn: Capoeta capoeta umbla) originating from Tigris River system (Kılıç-Demirok and Ünlü, 2001), Capoeta capoeta gracilis originating from Sefidroud and Shahroud Rivers (Pourali et al. 2006), Capoeta damascina originating from Ceyhan and Seyhan River system (Ünal, 2015) carry the same number of chromosomes with C. antalyensis and C. baliki. Besides, C. umbla bears significant similarities to C. antalyensis and C. baliki in terms of chromosome morphology. The only difference between them is that a chromosome pair identified as ST-A in C. antalyensis is M-SM in C. umbla and a chromosome pair identified as M-SM in C. baliki is ST-A in C. umbla. Also C. damascina is similar to C. baliki in terms of the number of M-SM and ST-A chromosome pairs whereas is different from C. antalyensis in terms of the number of chromosome Figure 1. Map shows the sampling sites.

3 M. K. Ayata et al. / Turk. J. Fish. Aquat. Sci. 17: (2017) 271 pairs classification as M-SM and/or ST-A. However, there are occasional differences between the chromosome morphologies of C. trutta on one hand and C. antalyensis and C. baliki on the other. C. antalyensis and C. baliki have a higher number of M- SM chromosome pairs and a lower number of ST-A chromosome pairs than C. trutta. Furthermore, number of arms of C. antalyensis and C. baliki is higher than C. trutta and C. umbla. Moreover C. baliki has the same number of arms with C. damascina but number of arms of C. antalyensis is lower than C. damascina. On the other hand, diploid chromosome number of C. antalyensis and C. baliki is the same with Capoeta capoeta (Safar et al., 2000) and Capoeta sevangi (Syn: Varicorhinus capoeta) (Krysanov, 1999) but it is different from C. damascina (Gorshkova et al., 2002). In terms of chromosome morphology C. antalyensis and C. baliki are very different from C. sevangi but they are very similar with the others. Moreover, number of arms of C. antalyensis is the same with C. capoeta. Otherwise Figure 2. (a) Metaphase and (b) karyotype of Capoeta antalyensis. Bar represents 3 µm.

4 272 M. K. Ayata et al. / Turk. J. Fish. Aquat. Sci. 17: (2017) number of arms of C. antalyensis and C. baliki is higher than C. sevangi but it is lower than C. damascina. Kılıç-Demirok and Ünlü (2001) reported that C. trutta and C. umbla could also be hexaploid species. Apart from cyprinids, Misgurnus angillicaudatus of the Cobitidae family was noted to be a hexaploid species having 6n=150 chromosomes (Abbas et al., 2009). Chromosome number of the hexaploid Carassius gibelio (Syn: Carassius auratus gibelio) was found 2n=160 (Mayr et al., 1986). These studies suggest that C. antalyensis and C. baliki may also be hexaploid species. Just like C. sevangi (Krysanov, 1999), C. trutta, C. umbla (Kılıç-Demirok and Ünlü, 2001) and C. damascina (Ünal, 2015) and as well as many other species in the same family (Rab and Collares-Pereira, 1995), C. antalyensis and C. baliki were also found to lack sex chromosome differentiation. Fishes show more extensive chromosomal diversity. Determination of numerical and structural chromosome differences are essential for genetic data Figure 3. (a) Metaphase and (b) karyotype of Capoeta baliki. Bar represents 3 µm.

5 M. K. Ayata et al. / Turk. J. Fish. Aquat. Sci. 17: (2017) 273 Table 1. Karyotype characteristics of Capoeta species that prevail in the inland waters of Turkey Species 2n Chromosome morphology NF References C. trutta M-SM+80ST-A 220 Kılıç-Demirok and Ünlü, 2001 C. umbla M-SM+64ST-A 236 Kılıç-Demirok and Ünlü, 2001 C. damascina M+42SM+62ST-A 238 Ünal, 2015 C. antalyensis M-SM+66ST-A 234 In this study C. baliki M-SM+62ST-A 238 In this study of species. It is believed that the results we have obtained will contribute to the cytogenetics of C. antalyensis and C. baliki. References Abbas, K., Li, M. Y., Wang, W. M. and Zhou, X. Y First record of the natural occurrence of hexaploid loach Misgurnus anguillicaudatus in Hubei Province, China. Journal of Fish Biology, 75: doi: /j x Collares-Pereira, M. J In vivo direct chromosome preparation (protocol for air drying technique). First International Workshop on Fish Cytogenetic Techniques, France, Gaffaroğlu, M., Karasu, M. and Unal, S Karyotype of river loach Turcinoemacheilus kosswigi Bănărescu and Nalbant, 1964 (Cypriniformes, Balitoridae) from the Euphrates River, Turkey. Journal of Agricultural Science and Technology, 14: Gaffaroğlu, M, Yüksel, E. and Rab, P Note on the karyotype and NOR phenotype of leuciscine fish Acanthobrama marmid (Osteichthyes, Cyprinidae). Biologia Bratislava, 61(2): doi: /s y Geldiay, R. and Balık, S Freshwater fish of Turkey. Ege University Press, İzmir. Gorshkova, G., Gorshkov, S. and Golani, D Karyotypes of Barbus canis and Capoeta damascina (Pisces, Cyprinidae) from the Middle East. Italian Journal of Zoology, 69: doi: / Guegan, J. F., Rab, P., Machordomo, A. and Doadrios, I New evidence of hexaploidy in 'large' African Barbus with some considerations on the origin of hexaploidy. Journal of Fish Biology, 47: doi: /j tb01888.x Kılıç-Demirok, N. and Ünlü, E Karyotypes of cyprinid fish Capoeta trutta and Capoeta capoeta umbla (Cyprinidae) from the Tigris River. Turkish Journal of Zoology, 25: Krysanov, E. Yu Karyotypes of Varicorhinus capoeta and Barbus goktschaicus (Cypriniformes) from lake Sevan, Armenia. Journal of Ichthyology, 39: ISSN Kuru, M., Yerli, S. V., Mangıt, F., Ünlü, E. and Alp, P Fish biodiversity in inland waters of Turkey. Journal of Academic Documents for Fisheries and Aquaculture, 3: Levan, A., Fredga, K. and Sandberg, A. A Nomenclature for centromeric position on chromosomes. Hereditas, 52: doi: /j tb01953.x Mayr, B., Rab, P. and Kalat, M NORs and counterstain-enhanced flourescence studies in Cyprinidae of different ploidy level. Genetica, 69: Oellerman, L. K. and Skelton, P. H Hexaploidy in yellowfish species (Barbus, Pisces; Cyprinidae) from Southern Africa. Journal of Fish Biology, 37: doi: /j tb05932.x Pourali, D. S., Bazyar, L. A. and Hasanzadeh, K. B A karyological study of Barbus capito, Barbus mursa and two populations of Capoeta capoeta from Northern Iran. Iranian Journal of Natural Resources, 58(4): Rab, P. and Collares-Pereira, M. J Chromosomes of European Cyprinid fishes (Cyprinidae, Cypriniformes): A review. Folia Zoologica, 44(3): Safar, P., Mahmood, K., Bahram, K. and Masoud, S Karyological study of two populations of Capoeta capoeta from North Iran. Cytologia, 65: Thorgaard, G. H. and Disney, J. E. (1990). Chromosome preparation and analysis. In: Schreck, C. B, Moyle, P. B. ed. Methods for fish biology, American Fisheries Society, Maryland, USA, 29pp. Ünal, S Seyhan ve Ceyhan nehir sisitemlerinde yaşayan bazı cyprinid türlerinde karyolojik araştırmalar. PhD thesis, Ahi Evran University, Kırşehir.